A SCREENING METHOD FOR OXYGEN-TOLERANT HYDROGENASE BY THE EXPRESSION OF CYANOBACTERIAL HYDROGENASE IN RHODOBACTER SPHAEROIDES MUTANT

摘要

PURPOSE: A method for screening oxygen tolerant hydrogenase using a Rhodobacter sphaeroides mutant is provided to screen the hydrogenase without measuring hydrogen generation amount. CONSTITUTION: A method for screening oxygen tolerant hydrogenase comprises: a step of preparing a Rhodobacter sphaeroides mutant in which quinine oxidase(QxtAB) gene and cytochrome bc1(FbcBC) gene are deleted; and a step of expressing foreign hydrogenase in the mutant strain and confirming the growth of the mutant strain. A method for producing hydrogen comprises a step of culturing recombinant Rhodobacter sphaeroides which is transformed using the recombinant vector under anaerobic and dark conditions. The recombinant vector contains a gene encoding the foreign hydrogenase. [Reference numerals] (AA) Successive removal of Quinol oxidase Qxt/Cytochromebcl; (BB) Introduction; (CC) Manufacturing a structure expressing cyanobacterium-derived hydrogenase; (DD) Acquiring recombinant; (EE) Separating suppressor mutant which survives under various oxygen partial pressures of 1%-20%; (FF) Acquring and analyzing HoxYH mutein